Objective:To establish an allele-specific polymerase chain reaction(PCR)identification method for vinegar Testudinis Carapax et Plastrum formula granules.Methods:Based on the single nucleotide polymorphism on the cytochrome C oxidase subunit Ⅰgene sequence of Chinemys reevesii(Gray)and its adulterant,specific primers of vinegar Testudinis Carapax et Plastrum were designed.The optimal PCR reaction system was determined by optimizing the reaction system,and the tolerability and applicability of this method were investigated.Results:When the annealing temperature was 62℃,and the cycle number was 34,the specific band of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules could be obtained at about 92 bp,while no band was found in other adulterants and negative control.Conclusion:The established allele-specific PCR identification method can quickly and accurately identify the authenticity of vinegar Testudinis Carapax et Plastrum decoction pieces,standard decoction freeze-dried powder,and formula granules,providing a reference for the research on quality standards of vinegar Testudinis Carapax et Plastrum.
关键词
醋龟甲/配方颗粒/位点特异性聚合酶链式反应/分子鉴定
Key words
vinegar Testudinis Carapax et Plastrum/formula granules/allele-specific PCR/molecular identification
维普
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